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Research ArticleEvaluation of Toxicity and Antimicrobial Activity ofan Ethanolic Extract from Leaves of Morus alba L. (Moraceae)

Alisson Macário de Oliveira,1,2 Matheus da Silva Mesquita,2

Gabriela Cavalcante da Silva,2 Edeltrudes de Oliveira Lima,3 Paloma Lys de Medeiros,4

Patrícia Maria Guedes Paiva,1 Ivone Antônia de Souza,2 and Thiago Henrique Napoleão1

1Departamento de Bioquımica, Centro de Ciencias Biologicas, Universidade Federal de Pernambuco, 50670-420 Recife, PE, Brazil2Laboratorio de Farmacologia e Cancerologia Experimental, Departamento de Antibioticos, Universidade Federal de Pernambuco,50670-420 Recife, PE, Brazil3Laboratorio deMicologia, Departamento de Ciencias Farmaceuticas, Centro de Ciencias da Saude, Universidade Federal da Paraıba,58059-900 Joao Pessoa, PB, Brazil4Departamento de Histologia e Embriologia, Centro de Ciencias Biologicas, Universidade Federal de Pernambuco,50670-420 Recife, PE, Brazil

Correspondence should be addressed toThiago Henrique Napoleao; [email protected]

Received 4 May 2015; Accepted 28 June 2015

Academic Editor: Ilaria Lampronti

Copyright © 2015 Alisson Macario de Oliveira et al. This is an open access article distributed under the Creative CommonsAttribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work isproperly cited.

This work evaluated an ethanolic extract from Morus alba leaves for toxicity to Artemia salina, oral toxicity to mice, andantimicrobial activity. Phytochemical analysis revealed the presence of coumarins, flavonoids, tannins, and triterpenes in theextract, which did not show toxicity to A. salina nauplii. No mortality and behavioral alterations were detected for mice treatedwith the extract (300 and 2000mg/kg b.w.) for 14 days. However, animals that received the highest dose showed reduced MCV andMCHC as well as increased serum alkaline phosphatase activity. In treatments with the extract at both 300 and 2000mg/kg, therewas a reduction in number of leukocytes, with decrease in percentage of lymphocytes and increase in proportion of segmentedcells. Histopathological analysis of organs frommice treated with the extract at 2000mg/kg revealed turgidity of contorted tubulesin kidneys, presence of leukocyte infiltration around the liver centrilobular vein, and high dispersion of the spleen white pulp. Theextract showed antimicrobial activity against Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, Candida krusei,Candida tropicalis, andAspergillus flavus. In conclusion, the extract contains antimicrobial agents and was not lethal for mice wheningested; however, its use requires caution because it promoted biochemical, hematological, and histopathological alterations.

1. Introduction

The use of plants as therapeutic tools is based on the popularculture and plant preparations are continually used by people,despite the large number of synthetic drugs developed. Thishas beenmainly associated with the higher costs of allopathicmedicines and a limited access of population to them,especially in developing countries where most of the peopledepend essentially on natural resources to address problemsrelated to diseases of primary care [1]. The relevance ofmedicinal plants over the centuries has stimulated the search

for scientifically proven information on their efficacy andsafety for humans [2–4].

Morus genus (Moraceae) comprises a group of treesnative from Asia, popularly known as mulberries, and witha great importance in folk medicine. In China, the leaves,roots, and branches of Morus species are used for treatmentof fevers, hepatic protection, vision improvement, strength-ening of joints, and reduction of blood pressure [5–7].The leaves of different mulberries are consumed in Koreaand Japan as a nutraceutical food and used for controllingblood glucose levels; this last property is attributed to the

Hindawi Publishing CorporationEvidence-Based Complementary and Alternative MedicineVolume 2015, Article ID 513978, 7 pageshttp://dx.doi.org/10.1155/2015/513978

2 Evidence-Based Complementary and Alternative Medicine

presence of the compound 1-deoxynojirimycin, a potent 𝛼-glucosidase inhibitor [8]. Fukai et al. [9] isolated fromMorusspecies a substance deemed chalcomoracin, which showedantimicrobial activity against methicillin-resistant Staphylo-coccus aureus. Dai et al. [10] isolated three compounds fromMorus macroura bark, called guangsangon H, guangsangonI, and guangsangon J, which showed anti-inflammatory andantioxidant activities.

The pharmacological potential of the species Morus albaL. (white mulberry), for benefit of animals and humans, hasbeen investigated. Flavonoids from root showed antiparasiticactivity against Ichthyophthirius multifiliis, a parasite of gillsand skin of freshwater fishes [11] and it was reported thata methanolic extract from the plant foliage might be usedas a dietary supplement in order to alleviate Aeromonashydrophila infection in catfish Clarias gariepinus [12]. Diels-Alder adducts and prenylated flavanones isolated from theroot bark of M. alba showed cytotoxic activity againsthuman tumor cells [13]. In another study, a compositioncontaining a blend of standardized extracts from Uncariagambir leaves and M. alba root bark was reported to beuseful as an alternative therapy for alleviating osteoarthritisand its associated symptoms [14]. Kim et al. [15] reportedthat extracts from M. alba leaves and fruits were able toreduce cognitive deficits in mice induced by the high-fat diet.Other reports mention antioxidant, antibacterial, antiviral,and neuroprotective activities from extracts and isolatedcompounds from fruits, leaves, stem, and root ofM. alba [16–20].

In face of the several reports on the medicinal propertiesof M. alba leaves, we investigated in this work the toxicityof an ethanolic extract from this tissue by using two models:Artemia salina lethality assay and assessment of in vivo oraltoxicity to mice. In the last, biochemical, hematological, andhistopathological analyses were also performed. In addition,it reports the phytochemical composition of the extract andthe evaluation of its antimicrobial activity against pathogenswith medical relevance.

2. Materials and Methods

2.1. Plant Material. Leaves of M. alba were collected inPetrolina city, Pernambuco, northeast Brazil. Taxonomicidentification was performed and a voucher specimen (num-ber 88372) is deposited in the herbarium of the InstitutoAgronomico de Pernambuco, Recife, Brazil.

2.2. Extract Preparation. The ethanolic extract was chosenfor this study because this solvent is able to dissolve bothpolar and nonpolar substances, including a variety of plant-derived compounds. The leaves were washed with distilledwater and dried at 28∘C during 24 h and next in an ovenat 45∘C for 15 days; subsequently, the leaves were powdered.The leaf powder (180 g) was mixed with 70% (v/v) ethanol indistilled water and the mixture was allowed to rest for 48 h,followed by mechanical agitation for 48 h using an orbitalshaker. The extract was filtered and the solvent was removedusing a rotary evaporator.

Table 1: Phytochemical screening of ethanolic extract from Morusalba leaves through thin layer chromatography.

Compound classes Revealer ResultAlkaloids Dragendorff ’s reagent NegativeAnthraquinones KOH NegativeCoumarins KOH Positive

Steroids Liebermann-Burchard Negative

Flavonoids Aluminum chloride Positive

Tannins Iron (III) chloride Positive forcondensed tannins

Triterpenes Liebermann-Burchard Positive

2.3. Phytochemical Screening. The extract was evaluated forthe presence of coumarins, flavonoids, tannins, steroids,alkaloids, anthraquinones, and triterpenes by thin layerchromatography (TLC). The assays were performed usingthe revealers listed in Table 1 and following the instructionsdescribed by Markhan [21], Wagner and Bladt [22], andAbreu [23].

2.4. Toxicity to Artemia salina. The assay was performedaccording to Meyer et al. [24] using A. salina cysts purchasedfrom San Francisco Bay Brand, Inc. (USA). First, an artificialsaline solution (3.8%, w/v) was prepared using sea salt(Marinex, Brazil) and distilled water. In each assay, 10 naupliiwere added to extract solutions at different concentrations(100–1,000𝜇g/mL) prepared by diluting the extract in 10mLof the artificial saline solution. In controls, the naupliiwere incubated with the saline solution. Each concentrationwas tested in triplicate and three independent experimentswere performed. The assays were maintained under artificiallighting for 24 h at 27 ± 2∘C. After this period, the mortalityrates were determined.

2.5. Oral Toxicity Assay. The experiments were performedusing female Swiss mice (Mus musculus) weighing 38–50 gand reared in the vivariumof theLaboratorio de Farmacologiae Cancerologia Experimental of the Universidade Federal dePernambuco. The animals are maintained at temperatureof 21 ± 1∘C, 12L : 12D photoperiod and it is given adlibitum access to food (Purina) and water. The experimentalprocedures were approved by Animal Ethics Committee ofthe Universidade Federal de Pernambuco (process number23076.061132/2014-33).

The dried extract was dissolved in 0.9% (w/v) NaCl [25]and acute toxicity (mortality and behavioral alterations) wasevaluated by oral administration. The mice were separatedin three groups (𝑛 = 3 for each group), according to theinstructions of the Organization for Economic Cooperationand Development [26]: control group, which received salinesolution (vehicle), and two test groups, which received theextract at 300mg/kg and 2000mg/kg b.w. The mice wereobserved during 14 days.

Evidence-Based Complementary and Alternative Medicine 3

2.6. Biochemical and Hematological Analyses. At the end oforal toxicity assays, the blood of animals was collected andthe following biochemical parameters were evaluated: totalprotein, albumin, alanine aminotransferase (ALT), aspar-tate aminotransferase (AST), alkaline phosphate, gamma-glutamyl transferase (GGT), urea, and creatinine, usingspecific kits (Labtest Diagnostica, Lagoa Santa, Brazil) anda COBAS Mira Plus analyzer (Roche Diagnostics Systems,Basel, Switzerland). Hematologic analysis was performedusing an automatic analyzer (Animal Blood Counter: ABCVet, Montpellier, France) and optical microscopy; the param-eters evaluated were as follows: erythrocytes, hemoglobin,hematocrit, mean corpuscular volume (MCV), mean cor-puscular hemoglobin (MCH),mean corpuscular hemoglobinconcentration (MCHC), and total and differentiated analysisof leukocytes.

2.7. Histopathological Analysis. Histological analyses of liver,kidney, and spleen of animals from control and extracttreatments were performed by opticalmicroscopy. Fragmentsof the organs were fixed in buffered formalin (10%, v/v)and then dehydrated through a graded ethanol series (70–100%), diaphonized in xylol, and embedded in paraffin.Histological sections (5 𝜇m) were stained with hematoxylin-eosin and mounted using cover slips with Entellan resin(Merck, Germany) [27].Thematerials were observed under aMotic BA200 microscopy coupled to a Moticam 1000 1.3MPdigital camera (Motic Incorporation Ltd., Causeway Bay,Hong Kong).

2.8. Antimicrobial Activity. The extract was evaluated forantibacterial activity against three strains of Staphylococcusaureus (ATCC-13150, M-177, and LM-197) and two strains ofPseudomonas aeruginosa (ATCC-9027 and P-03). Antifungalactivity was evaluated against strains of Candida albicans(ATCC-76645 and LM-106), Candida tropicalis (ATCC-13803 and LM-6), Candida krusei (LM-656 and LM-978),Aspergillus flavus (LM-118), and Aspergillus niger (LM-108).Bacterial suspensions were prepared inNutrient Broth (DifcoLaboratories, France) and fungal suspensions in RPMI 1640medium (Acumedia, India). The suspensions were standard-ized for 106 colony-forming units (CFU) permLusing a Leitz-Photometer 340–800 spectrophotometer.

The assays for determination of the minimal inhibitoryconcentrations (MIC) were performed in U-bottom 96-wellmicroplates. In an assay, each plate well received 100 𝜇L ofNutrient Broth (for bacteria) or RPMI medium (for yeastsand filamentous fungi). Next, 100𝜇L of the extract, dissolvedin distilled water, was added in the wells of the first columnof the plate and then a two-fold serial dilution was performedin each row to reach extract concentrations ranging from 32to 1,024 𝜇g/mL. Finally, 10𝜇L of microorganism suspensionwas added to each well. A 100% growth control was done inabsence of the extract and positive controls were performedusing chloramphenicol (100 𝜇g/mL) for bacteria as well asnystatin (100 IU) and fluconazole (100 𝜇g/mL) for fungi. Theplates were incubated for 24 h at 35∘C in assays with bacteriaand yeasts or for 7–10 days at 28∘C in assays with filamentous

fungi. After 24 h, bacterial growthwas revealed by addition of20𝜇L of 0.01% (w/v) resazurin (Inlab Confianca, Sao Paulo,Brazil); a change in color from blue to red was indicativeof microbial growth and thus the MIC was recorded as thelowest extract concentration at which there was no colorchange [28]. For the fungi, the MIC was defined as the lowestconcentration able to inhibit the visual growth in comparisonwith the 100% growth control. Each antimicrobial assaywas performed in duplicate and repeated three times. Theantimicrobial activity of the extract was classified accordingto MIC values as follows: 50–500𝜇g/mL = strong activity;600–1500𝜇g/mL = moderate activity; and >1500𝜇g/mL =weak activity or inactive [29].

2.9. Statistical Analysis. The data were expressed as mean ±SD and submitted to one-way ANOVA followed by Bonfer-roni’s test, with significance level at 𝑝 < 0.05.

3. Results and Discussion

Health treatments using medicinal plants are the less expen-sive and more accessible to the population, mainly in devel-oping countries. However, the indiscriminate use of plantpreparations or products with doubtful constitution andwithout rigorous standardization may pose a risk due to thetoxicity of some plant compounds to human organism. Forthis reason, studies that evaluated the side effects and toxicitydegree of preparations and compounds frommedicinal plantsare imperative. Within this context, this work evaluated thetoxicity of an ethanolic extract from leaves ofM. alba, a plantwhose tissues have been extensively used with medicinalpurposes, including as commercial formulations.

The phytochemical screening of the extract revealed thepresence of coumarins, flavonoids, tannins, and triterpenes(Table 1). Toxicity of the extract was firstly evaluated usingthe microcrustacean A. salina, which is often used as apreliminary indicator of general toxicity of plant compounds;also, it is reported that toxicity to A. salina has a correlationwith possible antitumor activity [30, 31]. The results showedthat the extract hadno toxicity at all the tested concentrations,suggesting that it does not have a general toxicity.

In the oral toxicity assays, the groups ofmice from controland extract treatments at both 300 and 2000mg/kg b.w.did not show alterations in behavioral signals during the 14days of experiments. However, the hematological analysis(Table 2) demonstrated significant (𝑝 < 0.05) reductionsin MCV and MCHC in animals treated with the extract at2000mg/kg, in comparison with control; MCHC was alsolower than control in treatment at 300mg/kg. This resultindicates that components of the M. alba extract may beacting on the erythrocytes causing a reduction of hemoglobincontent and may be interfering with the hematopoiesis,causing the production of cells with lower volume. It hasbeen reported that plant secondary metabolites may actdirectly on erythrocytes interfering with the integrity ofplasma membrane and then causing shrinkage, reduction inhemoglobin content, and even destruction of the cells [32].


Table 2: Hematological parameters of animals from control group and treated with ethanolic extract ofMorus alba leaves for 14 days.

Parameter Control Extract300mg/kg b.w. 2000mg/kg b.w.

Erythrocytes (106/mm3) 7.05 ± 0.39 7.84 ± 0.58 6.82 ± 0.51Hematocrit (%) 39.95 ± 0.21 41.47 ± 2.41 36.50 ± 4.95Hemoglobin (g/dL) 17.35 ± 1.20 15.62 ± 1.05 16.00 ± 1.41MCV (fL) 64.00 ± 5.00 64.33 ± 5.03Δ 50.00 ± 1.00∗Δ

MCH (pg) 26.97 ± 1.91 24.80 ± 0.85 21.20 ± 4.53MCHC (%) 38.20 ± 1.01 32.67 ± 0.85∗ 32.20 ± 2.01∗

Leukocytes (103/mm3) 8.26 ± 0.43 9.57 ± 0.66Δ 6.81 ± 0.27∗Δ

Segmented (%) 22.68 ± 1.16 43.90 ± 3.41∗Δ 55.71 ± 3.12∗Δ

Lymphocytes (%) 58.88 ± 1.70 42.39 ± 2.57∗Δ 29.65 ± 0.92∗Δ

Monocytes (%) 18.44 ± 3.55 13.71 ± 1.35 14.61 ± 0.64∗Significantly different (𝑝 < 0.05) from control treatment. ΔSignificantly different (𝑝 < 0.05) from the other dose of M. alba leaf extract. MCV: meancorpuscular volume; MCH: mean corpuscular hemoglobin; MCHC: mean corpuscular hemoglobin concentration.

Table 3: Biochemical parameters of blood of animals from control and treatments with the ethanolic extract fromMorus alba leaves for 14days.

Parameters Control Extract300mg/kg b.w. 2000mg/kg b.w.

Albumin (g/dL) 2.14 ± 0.17 1.92 ± 0.13 2.25 ± 0.25ALT (U/L) 78.50 ± 2.12 80.3 ± 8.02Δ 66.33 ± 4.73∗Δ

AST (U/L) 147.50 ± 4.95 149.67 ± 6.81 149.33 ± 5.03Total protein (g/dL) 6.16 ± 0.23 6.09 ± 0.35 6.13 ± 0.17Alkaline phosphatase (IU/L) 14.00 ± 1.0 12.33 ± 2.52Δ 37.00 ± 2.65∗Δ

GGT (U/L) 11.00 ± 1.41 12.67 ± 1.15 14.00 ± 1.50Urea (mg/dL) 54.33 ± 4.93 50.33 ± 1.53 53.33 ± 2.08Creatinine (mg/dL) 0.28 ± 0.02 0.28 ± 0.02 0.30 ± 0.04∗Significantly different (𝑝 < 0.05) from control treatment. ΔSignificantly different (𝑝 < 0.05) from the other dose ofM. alba leaf extract.

Table 2 also shows that the number of leukocytes in ani-mals treated with the extract at 2000mg/kg was significantlylower (𝑝 < 0.05) than in control animals, having observed areduction in the proportion of lymphocytes but an increase ofsegmented leukocytes. At the dose of 300mg/kg, the extractofM. alba also decreased the proportion of lymphocytes andincreased the percentage of segmented leukocytes, althoughthe number of total leukocytes was not altered. A possibleexplanation is that compounds of the extract resulted ina drastic decrease of blood lymphocytes, which stimulatedthe production of new leukocytes, leading to the increasein proportion of segmented cells. In summary, these resultsconstitute an indication that this extract has an importanteffect on the immune cells.

Biochemical analysis (Table 3) revealed significant (𝑝 <0.05) alterations in the serum levels of the enzymes ALT andalkaline phosphatase in animals that received the extract at2000mg/kg, in comparison with control. The group treatedwith the dose 300mg/kg did not show significant (𝑝 > 0.05)variations. Since damage of liver cells usually results in arelease of several types of enzymes, we can infer that the M.alba extract did not cause strong impairment of hepatocytesmembrane. Indeed, no alterations or a decrease only of ALTlevels, which is a more specific marker for damage to liver

Table 4: Evaluation of food andwater consumption andweight gainof animals from control and treated with the ethanolic extract fromleaves ofMorus alba.

Parameter Control Extract300mg/kg b.w. 2000mg/kg b.w.

Water consumed(mL) 35.00 ± 1.05 34.82 ± 1.68 34.29 ± 1.27

Food consumed(g) 28.68 ± 1.23 28.13 ± 1.00 27.09 ± 1.28

Weight gain (g) 12.04 ± 0.79 11.98 ± 0.49 12.00 ± 0.38

cells, has been interpreted as a signal of stability of themembranes of these cells [33]. An increase only in alkalinephosphatase level is usually indicative of obstruction of bileflow [4] and it is possible that theM. alba extract is acting atthis level and not by disturbing themembrane of hepatocytes.The levels of GGT, which is amore specificmarker for hepaticdamage [34], were similar in all groups, corroborating thattheM. alba extract did not exert hepatotoxic effect.

No differences in food and water consumption weredetected between the groups as well as regarding the biomassgain (Table 4). The results from oral toxicity evaluation


300mg/kg

2000mg/kg

KidneyCon

trol

Liver Spleen

Figure 1: Histopathological analysis of kidneys, livers, and spleens ofmice from control and treatments with the ethanolic extract fromMorusalba leaves at 300 and 2000mg/kg b.w. Kidneys: in control and treatment with 300mg/kg, the cortical region with the renal glomeruli (∗) andcontorted tubules (arrows) without alterations can be seen. In treatment with 2000mg/kg, a reduction of the subcapsular space and turgidcontorted tubules (arrows) is observed. Livers: photomicrography from control treatment shows the centrilobular vein (v) and a preservedorganization of the hepatocytes bundles. In the image from treatment with 300mg/kg, reversible-type cytoplasm vacuolizations (arrows) canbe seen while the photomicrograph of an animal that received 2000mg/kg of extract shows a lymphocyte infiltration around the centrilobularvein (white arrow). Spleens: in control, the well-delimited lymphatic nodes (Nd) can be noticed, while, in images for treatments with 300 and2000mg/kg, there is an expansion of lymphatic node (Nd) in the white pulp. All sections were stained with hematoxylin/eosin. Kidney andliver images: 400x. Spleen images: 100x.

indicated that the ethanolic extract from leaves of M. albahas low toxicity, since no death was observed even in assaysat 2000mg/kg dose [26]. However, the alterations foundin hematological and biochemical analyses stimulated us toperform a histopathological study of the livers, kidneys, andspleens of the animals, since damage to these organs may belinked to the changes detected [35].

Photomicrographs obtained in the histopathologicalanalysis are showed in Figure 1. The structure of the organswas preserved in control animals while a slight presence ofreversible-type vacuolization was observed in hepatocytes ofanimals treated with the extract at 300mg/kg. On the otherhand, all the organs from animals that received the extractat 2000mg/kg showed alterations. The kidneys showed adecreasing of the subcapsular space and turgidity of thecontorted tubules. In the livers, the presence of leukocyte

infiltration around the centrilobular vein was detected, whichcan be associated with intracellular lesions. A high dispersionof the spleen white pulp was also observed, which is a regionof lymphatic tissue containing B and T lymphocytes; this canexplain the reduction in the number of lymphocytes recordedin hematological analysis.

The ethanolic extract ofM. alba leaves can be consideredmore toxic than an ethanolic extract from Ocimum sanctumleaves, which did not promote mortality at 200, 600, and2000mg/kg in 14 days and did not affect water consumption,bodyweight, and hematological and biochemical parameters.In addition, there were no alterations in the structure of liver,kidney, and spleen ofmice treatedwith theO. sanctum extractat a dose of 800mg/kg [36]. On the other hand, a leaf extractfrom Lawsonia inermis showed higher toxicity than the M.alba extract since it promoted pathological changes in liver


Table 5: Minimal inhibitory concentrations (MIC) of ethanolicextract fromMorus alba leaves against bacteria and fungi.

Microbial strain MIC (𝜇g/mL)Staphylococcus aureus ATCC-13150 512Staphylococcus aureusM-177 1,024Pseudomonas aeruginosa ATCC-9027 1,024Pseudomonas aeruginosa ATCC-P-03 1,024Candida albicans ATCC-76645 1,024Candida albicans LM-106 256Candida tropicalis ATCC-13083 512Candida tropicalis LM-6 1,024Candida krusei LM-656 512Candida krusei LM-978 512Aspergillus flavus KM-714 512Aspergillus niger LM-108 NINI: no inhibition.

and kidney of rats at a dose of 1000mg/kg, with signals ofdegenerative/apoptotic changes [37].

Coumarins are known for their hepatotoxicity and havebeen reported to be toxic to mice and rats, including beingable to increase the incidence of adenomas and carcinomasin lungs of rats [38, 39]. Thus, this class of compounds maybe involved in the alterations promoted by M. alba extractin oral toxicity assays. On the other hand, flavonoids areknown to exert hepatoprotective and antioxidant effects [40]and thus may counterbalance the effects of potentially toxiccompounds in the extract.

Among the several applications described for Morusplants, there is the use for combating infections caused bybacteria and fungi. The increase in the number of microbialstrains resistant to the mostly used antibiotics has stimulatedthe search for new antimicrobial agents, especially those withnatural origin [41]. In this way, we evaluated the antimicrobialactivity of the ethanolic extract from M. alba leaves againstbacteria and fungi with medical relevance. The results aresummarized in Table 5 and show that the extract presenteda strong activity (MIC of 256𝜇g/mL) against C. albicans LM-106 and moderate antimicrobial activity on the other strains,except A. niger, which was not sensitive to the extract. Thebacterium most sensitive to the extract was S. aureus ATCC-13150. The strains C. albicans ATCC-76645, C. albicans LM-106, and C. krusei LM-656 were resistant to the positivecontrols used.

4. Conclusions

Ethanolic extract of M. alba leaves showed low oral toxicityto mice since no animal death was detected at a dose of2000mg/kg. However, the extract promoted biochemical,hematological, and histopathological alterations at this dose,which indicates that caution is required regarding its use.At 300mg/kg, the extract did not show toxicity or causeirreversible cellular damages but altered the proportion ofleukocyte types.The extract showedmainlymoderate activityagainst the microbial pathogens evaluated.

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper.

Acknowledgments

The authors express their gratitude to the Fundacao deAmparo a Ciencia e Tecnologia do Estado de Pernambuco(FACEPE; APQ-0108-2.08/14), theConselho Nacional de Des-envolvimento Cientıfico e Tecnologico (CNPq; 472546/2012-0; 446902/2014-4) and the Coordenacao de Aperfeicoamentode Pessoal de Nıvel Superior (CAPES) for financial support.Patrıcia Maria Guedes Paiva thanks CNPq for investigatorresearch grant. Alisson Macario de Oliveira thanks CAPESfor graduate scholarship.

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